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Modeling the Effect of pH and Temperature for Cellulases Immobilized on Enzymogel Nanoparticles

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Abstract

Production costs of cellulosic biofuels can be lowered if cellulases are recovered and reused using particulate carriers that can be extracted after biomass hydrolysis. Such enzyme recovery was recently demonstrated using enzymogel nanoparticles with grafted polymer brushes loaded with cellulases. In this work, cellulase (NS50013) and β-glucosidase (Novozyme 188) were immobilized on enzymogels made of poly(acrylic acid) polymer brushes grafted to the surface of silica nanoparticles. Response surface methodology was used to model effects of pH and temperature on hydrolysis and recovery of free and attached enzymes. Hydrolysis yields using both enzymogels and free cellulase and β-glucosidase were highest at the maximum temperature tested, 50 °C. The optimal pH for cellulase enzymogels and free enzyme was 5.0 and 4.4, respectively, while both free β-glucosidase and enzymogels had an optimal pH near 4.4. Highest hydrolysis sugar concentrations with cellulase and β-glucosidase enzymogels were 69 and 53 % of those with free enzymes, respectively. Enzyme recovery using enzymogels decreased with increasing pH, but cellulase recovery remained greater than 88 % throughout the operating range of pH values less than 5.0 and was greater than 95 % at pH values below 4.3. Recovery of β-glucosidase enzymogels was not affected by temperature and had little impact on cellulase recovery.

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Abbreviations

PAA:

Poly(acrylic acid)

IUPAC:

International Union of Pure and Applied Chemistry

RSM:

Response surface methodology

ANOVA:

Analysis of variance

HPLC:

High-performance liquid chromatography

DNS:

Dinitrosalicylic acid

BSA:

Bovine serum albumin

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Acknowledgments

Funding for this research was provided by the National Science Foundation (Arlington, VA) under grant numbers CBET 0966526 and CBET 0966574.

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Author notes

  1. Andrey Zakharchenko & Sergiy Minko

    Present address: Nanostructured Materials Laboratory, University of Georgia, Athens, GA, 30602, USA

Authors and Affiliations

  1. Department of Agricultural and Biosystems Engineering, North Dakota State University, Fargo, ND, 58108-6050, USA

    Ashani Samaratunga, Nurun Nahar & Scott W. Pryor

  2. Department of Coatings and Polymeric Materials, North Dakota State University, Fargo, ND, 58102, USA

    Olena Kudina & Andriy Voronov

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  1. Ashani Samaratunga
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Correspondence to Scott W. Pryor.

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Samaratunga, A., Kudina, O., Nahar, N. et al. Modeling the Effect of pH and Temperature for Cellulases Immobilized on Enzymogel Nanoparticles. Appl Biochem Biotechnol 176, 1114–1130 (2015). https://doi.org/10.1007/s12010-015-1633-z

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